Aquatic Toxicology 70 (2004) 63–81 Physiological action of dissolved organic matter in rainbow trout in the presence and absence of copper: Sodium uptake kinetics and unidirectional flux rates in hard and softwater Aline Y.O. Matsuo a,b, , Richard C. Playle c , Adalberto L. Val a , Chris M. Wood b a Laboratory of Ecophysiology and Molecular Evolution, Instituto Nacional de Pesquisas da Amazˆ onia (INPA), Alameda Cosme Ferreira, 1756 – Aleixo, 69083-000, Manaus, AM, Brazil b Department of Biology, McMaster University, 1280 Main Street West, Hamilton, ON, Canada L8S 4K1 c Department of Biology, Wilfrid Laurier University, Waterloo, ON, Canada N2L 3C5 Received 9 March 2004; received in revised form 6 July 2004; accepted 19 July 2004 Abstract We investigated the physiological effects of dissolved organic matter (DOM) on sodium (Na + ) transport in juvenile On- corhynchus mykiss (2.5 g) in the presence and absence of simultaneous acute exposure to copper (Cu 2+ ; 0, 70, and 300 gl -1 ). Trout were acclimated in either hardwater (1000 M Ca 2+ ) or softwater (100 M Ca 2+ ), and DOM was tested at approx- imately 8 mg C l -1 using a natural (NOM) and a commercial (AHA) source. Ion transport was evaluated based on kinetics estimates (maximum Na + uptake rates, J max ; substrate affinity, K m ) and unidirectional flux measurements (J in , J out , J net ). J max was higher and unidirectional flux rates were greater in softwater-acclimated trout. Fish exposed to DOM alone in hardwater exhibited an increased Na + transport capacity indicated by both the kinetics (67% higher J max for AHA) and J in measurements (153% higher for AHA and 125% higher for NOM). In softwater, the effects of DOM alone on kinetic parameters and unidirec- tional flux rates were negligible. Cu 2+ affected Na + uptake by a mixed-type inhibition (both non-competitive and competitive). In hardwater, only K m was increased (i.e., affinity decreased), whereas in softwater, K m was increased and J max was decreased, with more marked effects at the higher Cu 2+ level. In hardwater, the stimulatory effect of AHA on J max persisted even in the presence of 300 gl -1 Cu 2+ , whereas both AHA and NOM prevented the increase in K m caused by Cu 2+ ; these effects were reflected in J in measurements. In softwater, AHA helped to protect against the increased K m caused by high Cu 2+ , but there was no protection against the inhibition of J max . Unidirectional flux measurements indicated that in softwater, Cu 2+ inhibited J in at 70 gl -1 , whereas at 300 gl -1 Cu 2+ , J out was also stimulated. Fish were more affected by Cu 2+ in softwater, as indicated by the inability to control diffusive losses of Na + and a reduced ability to take up Na + , but in the presence of DOM, losses were © 2004 Elsevier B.V. All rights reserved. Corresponding author. Tel.: +55 92 643 3187; fax: +55 92 643 3186. E-mail address: matsuoaline@aol.com (A.Y.O. Matsuo). 0166-445X/$ – see front matter © 2004 Elsevier B.V. All rights reserved. doi:10.1016/j.aquatox.2004.07.005